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Background    In situ  vaccination-induced local inflammatory response resulted in the establishment of a pool of tissue-resident memory T (T RM ) cells and new vessels after the resolution of inflammation. T RM  cells have received increasing attention; however, the role of new vessels in protective response is still unknown.Materials and Methods   We performed the laparotomy to access the stomach and injected alum-based vaccine into the gastric subserous layer (GSL). At 28 days post vaccination, a parabiosis mouse model along with depletion of anti-CD90.2 antibody was employed to explore the function of perivascular lymphocyte clusters in recall responses. The composition of the gastric lymphocyte clusters was analyzed by immunofluorescence staining. Antibody responses were detected using ELISA. Gastric lymphocytes were analyzed using flow cytometry.Results   GSL vaccination induced the formation of new vessels in the inflamed region. These new vessels were different from native vessels in that they were generally accompanied by perivascular lymphocyte clusters that mainly consisted of CD90-expressing cells. Additionally, histological analysis revealed the presence of CD4 +  and CD8 +  T cells in the perivascular lymphocyte clusters. Administration of a dose of an anti-CD90.2 antibody to GSL-vaccinated mice resolved these clusters. The efficacy of protection was compared in the parabiosis mice. Upon challenge, the presence of perivascular lymphocyte clusters was responsible for the fast recall response, as depletion of these clusters by CD90.2 antibody administration resulted in decreased expressions of VCAM-1, Madcam-1, and TNF- α , as well as lower recruitment of proinflammatory immune cells, decreased antibody levels, and poor protection.Conclusions   Our research demonstrates that  in situ  vaccination-induced regional inflammatory response contributes to optimal recall response not only by establishing a CD4 +  T RM  pool but also by creating an “expressway,” i.e., perivascular lymphocyte cluster.

journal of immunology research

Perivascular Lymphocyte Clusters Induced by Gastric Subserous Layer Vaccination Mediate Optimal Immunity against Helicobacter through Facilitating Immune Cell Infiltration and Local Antibody Response